• 제목/요약/키워드: NAD(P)H

검색결과 191건 처리시간 0.031초

Protective effects of 5-aminolevulinic acid on heat stress in bovine mammary epithelial cells

  • Islam, Md Aminul;Noguchi, Yoko;Taniguchi, Shin;Yonekura, Shinichi
    • Animal Bioscience
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    • 제34권6호
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    • pp.1006-1013
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    • 2021
  • Objective: Cells have increased susceptibility to activation of apoptosis when suffering heat stress (HS). An effective supplementation strategy to mimic heat-induced apoptosis of bovine mammary epithelial cells (MECs) is necessary to maintain optimal milk production. This study aimed to investigate possible protective effects of the anti-apoptotic activity of 5-aminolevulinic acid (5-ALA) against HS-induced damage of bovine MECs. Methods: Bovine MECs were pretreated with or without 5-ALA at concentrations of 10, 100, and 500 µM for 24 h followed by HS (42.5℃ for 24 h and 48 h). Cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Real-time quantitative polymerase chain reaction and Western blotting were used to explore the regulation of genes associated with apoptosis, oxidative stress, and endoplasmic reticulum (ER) stress genes. Results: We found that 5-ALA induces cytoprotection via inhibition of apoptosis markers after HS-induced damage. Pretreatment of bovine MECs with 5-ALA resulted in dramatic upregulation of mRNA for nuclear factor erythroid-derived 2-like factor 2, heme oxygenase-1, and NAD(P)H quinone oxidoreductase 1, all of which are antioxidant stress genes. Moreover, 5-ALA pretreatment significantly suppressed HS-induced ER stress-associated markers, glucose-regulated protein 78, and C/EBP homologous protein expression levels. Conclusion: 5-ALA can ameliorate the ER stress in heat stressed bovine MEC via enhancing the expression of antioxidant gene.

가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성 (Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus)

  • 박은미;염정주
    • 생명과학회지
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    • 제20권2호
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    • pp.260-268
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    • 2010
  • 가물치(Channa argus) 조직의 젖산탈수소효소 동위효소(EC 1.1.1.27, lactate dehydrogenase, LDH)를 정제하고 생화학적, 면역학적 및 역학적 방법으로 특성을 연구하였다. LDH 활성은 골격근이 380.4 units로 가장 높고 심장 13.4, 눈 3.5, 뇌 조직 5.4 units이었으며, 심장의 CS 활성은 20.7 unit로 가장 높고, LDH/CS는 골격근 172.9, 심장 0.6, 눈 0.32, 뇌 0.47이고, 단백질 양은 골격근 14.7 mg/g이며, 특이활성(units/mg)은 골격근 25.88, 심장 0.79, 눈 0.31, 뇌 1.38 units/mg이었으므로 골격근은 혐기적이고, 심장은 호기적이었다. LDH $A_4$, $B_4$, eye-specific $C_4$에 대한 항혈청을 사용한 Western blot, 면역침강반응 및 native-polyacrylamide gel electrophoresis에 의해 $A_4$, $A_3B$, $A_2B_2$, $AB_3$$B_4$가 모든 조직에서 확인되었고, 눈 조직에서 $C_4$$AC_3$, $A_2C_2$, $AC_3$, 뇌 조직에서 $A_3C$도 확인되었다. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ 동위효소는 affinity chromatography와 Preparative PAGE Cell에 의해 정제되었다. LDH $A_4$ 동위효소는 $NAD^+$ 유입 후 정제되었고, eye-specific $C_4$$A_4$에 이어 용출되기 시작하였으며 $B_4$는 buffer 유입 후 용출되었다. 정제한 결과 $A_4$$B_4$ 및 eye-specific $C_4$와 분자구조의 일부가 유사하였지만 $B_4$$C_4$는 서로 다른 것으로 나타났으므로, 하부단위체 A는 보존적이고, 하부단위체 B는 A보다 더 빠르게 진화된 것으로 보인다. 피루브산 10 mM에서 $A_4$ 동위효소 39.98%, $A_2B_2$ 21.28%, $B_4$ 19.67% 및 eye-specific $C_4$ 16.87%의 활성이 남아있었고, 피루브산에 대한 $Km^{PYR}$$A_4$ 0.17 mM, $B_4$ 0.27 mM, eye-specific $C_4$ 0.133 mM였다. $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$$AB_3$의 최적 pH는 각각 pH 6.50, pH 8.5, pH 5.5, pH 6.0-6.5, 5.0 및 pH 7.5였고, 동질사량체 $A_4$와 이질사량체 동위효소들은 넓은 pH 영역에서 안정하였다. 특히 골격근은 LDH 활성이 크므로 활동성이 크며, 눈조직에서 피루브산 친화력이 강한 eye-specific $C_4$에 의해 피루브산 대사가 빠르게 일어나고, 이어서 $A_4$에 의해 젖산이 산화되어지는 것으로 사료되므로, 종의 생태환경 및 먹이 획득 양식에 따라 LDH-C 발현, 기질에 대한 친화도 및 대사 시간이 다른 것으로 사료된다.

The impaired redox status and activated nuclear factor-erythroid 2-related factor 2/antioxidant response element pathway in wooden breast myopathy in broiler chickens

  • Pan, Xiaona;Zhang, Lin;Xing, Tong;Li, Jiaolong;Gao, Feng
    • Animal Bioscience
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    • 제34권4호
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    • pp.652-661
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    • 2021
  • Objective: Wooden breast (WB) is a novel myopathy affecting modern broiler chickens, which causes substantial economic losses in the poultry industry. The objective of this study was to evaluate the effect of WB abnormality on meat quality, redox status, as well as the expression of genes of the nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. Methods: A total of 80 broilers (Ross 308, 42 days of age, about 2.6 kg body weight) raised at Jiujin farm (Suqian, Jiangsu, China) were used. Twelve unaffected (no detectable hardness of the breast area) and twelve WB-affected (diffuse remarkable hardness in the breast muscle) birds were selected from the commercial broiler farm according to the criteria proposed by previous studies. Results: The results indicated that WB showed histological lesions characterized by fiber degeneration and fibrosis, along with an increase of muscle fiber diameter (p<0.05). Moreover, higher pH value, lightness, yellowness, drip loss and cooking loss were observed in the WB group (p<0.05). Compared with the normal breast (NOR) group, the WB group showed higher formation of reactive oxygen species (p<0.05), increased level of oxidation products and antioxidant activities (p<0.05), accompanied with mitochondrial damages and lower mitochondrial membrane potential (p<0.05). Meanwhile, the relative mRNA expressions of Nrf2 and its downstream antioxidant genes including heme oxygenase-1, NAD(P)H qui none dehydrogenase 1, glutathione peroxidase, superoxide dismutase, and glutamate-cysteine ligase were higher than those of the NOR group (p<0.05). Conclusion: In conclusion, WB myopathy impairs meat quality by causing oxidative damages and mitochondrial dysfunction in broilers, even though the activated Nrf2/antioxidant response element pathway provides protection for the birds.

알콜증류폐액을 이용한 빵효모배양에서 포도당, 암모늄 및 인산의 자동첨가에 의한 증균 (Increasing Cell Concentration by the Automatic Addition of Glucose, Ammonium and Phosphate in the Cultivation of a Baker′s Yeast in Alcohol Distillery Wastewater)

  • 이형춘
    • KSBB Journal
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    • 제18권3호
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    • pp.197-201
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    • 2003
  • 알콜증류 폐액에 빵효모를 배양하는 중에 포도당, 암모늄 및 인산을 자동 첨가하여 균체량을 효율적으로 증가시킴으로써 폐액의 재이용성을 향상시키고자 하였다. 포도당은 DO를 제어파라미터로 하는 제어방법을 사용하여 자동 첨가함으로서 그 농도를 300 mg/L 이하로 유지하였다. 암모늄은 pH-stat 방법을 사용하여 N $H_4$OH로서 자동 첨가함으로서 12.6~17.4mM의 낮은 농도범위로 유지하였다. 인산의 경우에는 분석방법으로써 ascorbic acid법을 채택한 자동화 FIA sysetm을 제작하여 사용하였다. 이 system으로 배양 19.4 시간 후부터 배양액을 자동분석하고 제어한 결과 배양액의 P $O_4$-P 농도를 23.3~43.4 mg/L 의 낮은 농도범위로 유지할 수 있었다. 세가지 영양성분을 자동첨가하면서 배양한 결과 증식의 정체없이 균체량을 33.0배 더 증가시킬 수 있었다. 비증식속도는 0.19 $hr^{-l}$였다.

Reduction of Azobenzene by Purified Bovine Liver Quinone Reductase

  • Kim, Kyung-Soon;Shin, Hae-Yong
    • BMB Reports
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    • 제33권4호
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    • pp.321-325
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    • 2000
  • Quinone reductase was purified to homogeneity from bovine liver by using ammonium sulfate fractionation, ionexchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene. The apparent Km for 1,4-benzoquinone and azobenzene was 1.64 mM and 0.524 mM, respectively. The reduction of azobenzene by quinone reductase was almost entirely inhibited by dicumarol or Cibacron blue 3GA, potent inhibitors of the mammalian quinone reductase. In the presence of 1.0${\mu}M$ Cibacron blue 3GA, azoreductase activity was lowered by 45%, and almost complete inhibition was seen above 2.0 ${\mu}M$ Cibacron blue 3GA.

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Lovastatin biosynthesis enhanced by thiamine in Aspergillus terreus

  • 안우석;한규범
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2002년도 생물공학의 동향 (X)
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    • pp.184-187
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    • 2002
  • Lovastatin is a cholesterol-lowering agent, which plays a role of an inhibitor of 3-hydroxy-3- methylglutaryl coenzyme A reductase (HMG-CoA). When thiamine was supplemented in 3L batch fermentation, the production of lovastatin was improved. At the same time, the levels of pyruvic acid and NAD(P)H were estimated in the course of the fermentation of A. terreus. For the high level production of lovastatin, semi fed-batch fermentation was performed. And the thiamine level was maintained to a concentration of 20 mg/L and glucose was supplied. The final dry cell weight was lowered by 30 % and final lovastatin concentration was increased by 33 %. Final lovastatin concentration of 3.3 g/L was achieved in 8 days.

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Occurrence of Thioredoxin Reductase in Deinococcus Species, the UV resistant Bacteria

  • Seo Hee-Jeong;Lee Young-Nam
    • Journal of Microbiology
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    • 제44권4호
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    • pp.461-465
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    • 2006
  • The occurrence of thioredoxin reductase (NAD(P)H: oxidized-thioredoxin reductase, EC 1.6.4.5, TrxR) in five mesophilic species of Deinococcus was investigated by PAGE. Each species possessed a unique TrxR pattern, for example, a single TrxR characterized D. radiopugnans while multiple forms of TrxR occurred in other Deinococcal spp. Most of TrxRs occurring in Deinococcus showed dual cofactor specificity, active with either NADH or NADPH, although the NADPH specific-TrxR was observed in D. radiophilus and D. proteolytic us.

Induction of Quinone Reductase , an Anticarcinogenic Marker Enzyme, by Vitamin E in Both Hepalclc7 Cells and Mice

  • Kwon, Chong-Suk;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • 제4권2호
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    • pp.122-124
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    • 1999
  • Induction of NAD(P)H : (quinone-acceptor) oxidoreductase (QR) which obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. We postulated that vitamin E, an antioxidant, which induces QR as the gene of QR was reported to contain antioxidant reponsive element in the 5'-flanking region. Vitamin E resulted in significant induction of QR in both hepalclc7 cells and mouse tissues. QR induction was observed; to be maximal at 25uM vitamin E for hepalclc7 cells while it was maximal in the level of 2.5∼5 μmoles vitamin E/㎏ BW for mouse tissues. Thus the cancer-preventive effect of vitamin E may be exerted by it induction of intracellular detoxifying enzymes.

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저품위 동광석의 세균침출에 관한 연구 1 (Studies on the Bacterial Leaching of Low-grada Copper Mlnerals)

  • 이강순;오재현;민봉희
    • 미생물학회지
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    • 제9권3호
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    • pp.121-129
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    • 1971
  • The purpose of this study was to investigate the proprieties of practical application and possibility of bacterial leaching method to the low copper minerals in Korea. Therfore, ore samples and mine water of the Talsung, Ilkwang, Taekwang, Yongho and Siheung were analyzed chemically, mineralogically, and bacteriologically. The results obtained were as follows ; 1. The range of pH value was 2.6-3.8. 2. Copper contents in ore samples of Talsung and Ilkwang were 0.38% and 0.63%, and those in mine water of Talsung nad ilkwang were 584 .$\mu$g and 15.$\mu$g per ml, respectively. 3. Ferric sulfate contents in mine water of Tlsung and Ilkwang were 27.2.$\mu$g and 10.5.$\mu$g per ml, respectively. 4. Ferrobacillus ferroosxidans was isolated and identified from Talsung and Ilkwang mine water and specially Thiobacillus thiooxidans was isolated from Ilkwang mine water.

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